Applitech F.SIGHT 2.0 Single-Cell Cloning and Printing System

Overview

The Applitech F.SIGHT 2.0 Single-Cell Cloning and Printing System is an engineered solution for high-integrity monoclonal cell line development, built upon Cytena’s proprietary Single-Cell Printing (SCP) technology — a validated, microfluidic inkjet-based dispensing platform compliant with regulatory expectations for clonal derivation. Unlike conventional limiting dilution or FACS-based approaches, the F.SIGHT 2.0 enables deterministic, image-verified deposition of individual viable cells directly into standard 96-well or 384-well plates under sterile conditions. Its core principle relies on piezoelectric-driven droplet generation, where each nanoliter-scale droplet contains one cell, precisely directed to predefined well coordinates via real-time vision-guided alignment. This ensures physical separation without shear stress-induced damage, preserving cellular viability and functional phenotype — critical for therapeutic antibody production, viral vector manufacturing, and regenerative medicine applications.

Key Features

• Image-validated single-cell deposition: Integrated high-resolution brightfield and optional fluorescence imaging captures pre- and post-printing frames for every well, enabling unambiguous traceability of clonal origin.
• Ultra-fast plate filling: Completes single-cell sorting in ≤2 minutes for 96-well plates and ≤8 minutes for 384-well plates — significantly accelerating clone screening timelines.
• Sterile, disposable EASY.ON cartridge: Pre-sterilized, single-use microfluidic print head eliminates cross-contamination risk and removes need for cleaning validation or carryover assessment.
• Cell focusing module: Actively positions target cells at the nozzle centerline prior to ejection, improving capture efficiency for low-abundance or fragile cell populations (e.g., primary T cells, iPSCs).
• Precision droplet targeting: Sub-50 µm positional accuracy ensures consistent droplet landing at the geometric center of each well bottom, optimizing post-sort imaging quality and reducing edge effects during colony expansion.
• Biosafety cabinet-compatible footprint: Compact dimensions (W × D × H: 360 × 420 × 320 mm) allow full operation inside Class II A2 laminar flow hoods without compromising sterility or workflow integration.

Sample Compatibility & Compliance

The F.SIGHT 2.0 supports broad cell type compatibility, including CHO-K1, HEK293, hybridomas, human PBMCs, CAR-T cells, iPSCs, and adherent or suspension lines with diameters ranging from 8–30 µm. It operates within ISO 5 (Class 100) environments when used inside certified biosafety cabinets. All image data — including timestamped, plate-coordinate-tagged microscopy frames — are stored locally in DICOM-compliant format with immutable metadata, supporting audit readiness for FDA 21 CFR Part 11, EMA Annex 11, and ICH Q5D requirements. The system meets GLP/GMP-aligned documentation standards for clonality assurance in biologics process development.

Software & Data Management

F.SIGHT Control Software v3.x provides intuitive, wizard-driven operation with configurable gating based on morphological parameters (diameter, circularity, aspect ratio) or fluorescence intensity (via integrated LED excitation). Every sorted event triggers synchronized image acquisition, automatic file naming (e.g., “A01_20240517_142231_BF_pre.jpg”), and time-stamped log export (.csv). Raw images and metadata are archived in a structured local repository with role-based access control. Export options include TIFF/OME-TIFF for third-party analysis and PDF reports compliant with internal SOPs. The software architecture supports API-based integration with LIMS and robotic platforms (e.g., Hamilton STAR, Tecan Fluent) via RESTful endpoints.

Applications

• Monoclonal antibody cell line development requiring ICH Q5D-compliant clonality documentation
• AAV and lentiviral vector producer cell line isolation for gene therapy programs
• CRISPR-edited clone isolation with phenotypic validation (e.g., knockout confirmation via fluorescent reporter)
• Stem cell subcloning with preservation of pluripotency markers post-sort
• Exosome-secreting subpopulation enrichment using membrane-dye or cargo-specific fluorescent probes
• High-throughput single-cell RNA-seq library prep with guaranteed input cell identity

FAQ

Does the F.SIGHT 2.0 require user calibration before each run?

No. The system performs automated nozzle health check, droplet trajectory verification, and focus calibration during startup — no manual intervention required.
Can the EASY.ON cartridge be reused after sterilization?

No. Cartridges are designed for single-use only and must be discarded after completion of a sorting session to maintain sterility and prevent carryover.
Is fluorescence capability standard or optional?

Fluorescence imaging is an optional hardware module; base configuration includes high-resolution brightfield only.
How is clonality evidence archived and retrieved for regulatory submission?

All images are stored with embedded plate coordinate metadata and SHA-256 checksums; export packages include audit trails, instrument logs, and PDF summary reports compliant with ALCOA+ principles.
What plate formats are supported beyond 96- and 384-well?

Only ANSI/SLAS-standard 96-well and 384-well plates with flat-bottom, tissue-culture-treated surfaces are validated and supported.